purification of periplasmic proteins with polymyxin

[marilyn yoder]

	i'm sorta desperate at the moment and wonder if anyone
	can give me any assistance,

	we are purifying a protein from the periplasmic space of
	E. coli.  we are trying to implement a procedure developed
	by Lehrer et al. in which the outer membrane is permeabilized with
	Polymyxin B.  the reference we have (Martin et al, JMB (1993) 230:1097)
	states to add '25 ml of buffer A containing 1 mg of Polymyxin B/ml
	to the cell pellet'.  there is no indication as to the quantity
	of cells that they had.  if anyone is familiar with this procedure,
	could you please send us some information regarding how many mg
	of polymyxin B to add to how many grams of cells??

	thanks in advance,

	marilyn yoder



-- 
Marilyn Yoder					phone:	816-235-1986
School of Biological Sciences			fax:	816-235-5158
Division of Cell Biology and Biophysics
University of Missouri-Kansas City		e-mail	myoder at vax1.umkc.edu
5100 Rockhill Rd
Kansas City,  MO  64110-2499